Interaction Details


Details of experiments between P31947 and Q9UQC2
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 128-137
Site: 14-3-3_pSer133
Modification: (SEP)133
Sequence: LRNVSSAGHG
Construct: LRNVS(pSer)AGHG (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 142
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 2

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 154-163
Site: 14-3-3_pSer159
Modification: (SEP)159
Sequence: RERKSSAPSH
Construct: RERKS(pSer)APSH (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 113
P value: 0.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 3

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 154-163
Site: 14-3-3_pSer159
Modification: (SEP)159
Sequence: RERKSSAPSH
Construct: RERKS(pSer)APSH (crude synthetic peptide)
Average holdup BI: -0.22
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 111
P value: 0.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 4

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 205-214
Site: 14-3-3_pSer210
Modification: (SEP)210
Sequence: NARSASFSQG
Construct: NARSA(pSer)FSQG (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 4.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.42
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 5

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 205-214
Site: 14-3-3_pSer210
Modification: (SEP)210
Sequence: NARSASFSQG
Construct: NARSA(pSer)FSQG (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 3.99
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 205-214
Site: 14-3-3_pSer210
Modification: (SEP)210
Sequence: NARSASFSQG
Construct: NARSA(pSer)FSQG (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 111
P value: 1.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: biotin-ttds-RSASF(pSer)QGTR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 211
Standard deviation of dissociation constant (10-6M): 16
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-09
Number of measurements: 3
Measured pKd: 3.68
DOI: 10.1101/2022.12.08.519103

Experiment series 8

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: RSASF(pSer)QGTR (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.43
Immobilized partner concentration (10-6M): 113
P value: 0.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 9

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: RSASF(pSer)QGTR (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 142
P value: 1.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: RSASF(pSer)QGTR (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.28
Immobilized partner concentration (10-6M): 111
P value: 0.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 11

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 259-268
Site: 14-3-3_pSer264
Modification: (SEP)264
Sequence: TEFRDSTYDL
Construct: TEFRD(pSer)TYDL (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
P value: 0.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 12

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 259-268
Site: 14-3-3_pSer264
Modification: (SEP)264
Sequence: TEFRDSTYDL
Construct: TEFRD(pSer)TYDL (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 0.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 13

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 259-268
Site: 14-3-3_pSer264
Modification: (SEP)264
Sequence: TEFRDSTYDL
Construct: TEFRD(pSer)TYDL (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.56
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 14

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 317-326
Site: 14-3-3_pSer322
Modification: (SEP)322
Sequence: PATPLSAYQI
Construct: PATPL(pSer)AYQI (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 1.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 15

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 317-326
Site: 14-3-3_pSer322
Modification: (SEP)322
Sequence: PATPLSAYQI
Construct: PATPL(pSer)AYQI (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 0.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 16

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 317-326
Site: 14-3-3_pSer322
Modification: (SEP)322
Sequence: PATPLSAYQI
Construct: PATPL(pSer)AYQI (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 2.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 17

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 467-476
Site: 14-3-3_pSer472
Modification: (SEP)472
Sequence: RAGDNSQSVY
Construct: RAGDN(pSer)QSVY (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 1.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 18

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 467-476
Site: 14-3-3_pSer472
Modification: (SEP)472
Sequence: RAGDNSQSVY
Construct: RAGDN(pSer)QSVY (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 19

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 467-476
Site: 14-3-3_pSer472
Modification: (SEP)472
Sequence: RAGDNSQSVY
Construct: RAGDN(pSer)QSVY (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 0.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 20

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 380-389
Site: 14-3-3_pThr385
Modification: (TPO)385
Sequence: RSVAATIPRR
Construct: RSVAA(pThr)IPRR (crude synthetic peptide)
Average holdup BI: 0.58
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 5.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.09
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 21

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 380-389
Site: 14-3-3_pThr385
Modification: (TPO)385
Sequence: RSVAATIPRR
Construct: RSVAA(pThr)IPRR (crude synthetic peptide)
Average holdup BI: 0.62
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 7.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.06
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 22

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 380-389
Site: 14-3-3_pThr385
Modification: (TPO)385
Sequence: RSVAATIPRR
Construct: RSVAA(pThr)IPRR (crude synthetic peptide)
Average holdup BI: 0.67
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 4.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.26
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 23

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 386-395
Site: 14-3-3_pThr391
Modification: (TPO)391
Sequence: IPRRNTLPAM
Construct: IPRRN(pThr)LPAM (crude synthetic peptide)
Average holdup BI: 0.85
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 5.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.69
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 24

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 386-395
Site: 14-3-3_pThr391
Modification: (TPO)391
Sequence: IPRRNTLPAM
Construct: IPRRN(pThr)LPAM (crude synthetic peptide)
Average holdup BI: 0.85
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 7.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.6
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 25

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: GAB2
Fragment: 386-395
Site: 14-3-3_pThr391
Modification: (TPO)391
Sequence: IPRRNTLPAM
Construct: IPRRN(pThr)LPAM (crude synthetic peptide)
Average holdup BI: 0.85
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 7.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.7
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))